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Biomedical subjects

S Hiraga

Publications and source records attributed to S Hiraga.

At least 55 records · Page 3Linked to original sources

T-cell clonal change after allo-kidney transplantation in humans.

Whether T cells circulating peripherally express changes at a clonal level after renal transplantation is uncertain. To clarify this issue, we analyzed T-cell clonality of peripheral blood lymphocytes (PBLs) in 12 renal transplant recipients by a novel polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) method that can discriminate T-cell clones with different T-cell receptor (TCR) Vbeta motifs. The PCR-SSCP study showed that after transplantation, only a few distinct T-cell clonotypes accumulated in the absence of clinical episodes, irrespective of the compatibility of HLA antigens. In contrast, various T-cell clones appeared in cases of acute rejection (AR) and infection. These subsided immediately after the AR was resolved; however, they remained long after the resolution of the infection. In a case of AR followed by an infectious episode, distinct T-cell clones appeared concomitantly with each episode. Several of them disappeared or remained thereafter. In one case, significant numbers of accumulating bands were observed by in-vitro stimulation by mixed lymphocyte reaction (MLR); several were identical to those found in vivo. However, some of those that did not appear in vitro were apparent in vivo. In conclusion, the appearance of T-cell clonotypes at a peripheral level indicates the existence of immunologically activated T-cell clones, which were significantly affected by immunosuppressive therapy. It was also determined that the T-cell immune system is much more complicated in vivo than in vitro.

Abscess↗

Antigenic and functional analyses of glycoprotein of rabies virus using monoclonal antibodies.

Thirty-five monoclonal antibodies (MAbs) against glycoprotein (G protein) of the RC-HL strain of the rabies virus have been established. Using these MAbs, two antigenic sites (I and II) were delineated on the G protein of the RC-HL strain in a competitive binding assay. Of these, 34 MAbs recognized the epitopes on site II. Site II was further categorized into 10 subsites according to their patterns in a competitive binding assay. Each site II-specific MAb showed 5 to 23 nonreciprocal competitions. The reactivities of 35 MAbs to rabies and rabies-related viruses in an indirect immunofluorescent antibody test showed that six MAbs in group A binded to rabies and rabies-related viruses and eight MAbs in group E reacted only with rabies viruses, considering that the former represent the genus-specific of Lyssavirus and the latter are rabies virus-specific. From biological assays, 28 of the 35 MAbs showed neutralization activity, 31 showed hemagglutination inhibition (HI) activity, and 18 showed immunolysis (IL) activity. The MAbs recognizing neutralization epitopes fell into at least three groups: those exhibiting both HI and IL activity, those showing only HI activity, and those showing neither HI nor IL activity. All IL epitopes overlap with HA epitopes. Five of the nine MAbs which reacted with the antigen treated by sodium dodecyl sulfate in ELISA were not reduced, or reduced only slightly, in the titer. None of the MAbs reacted with 2-mercaptoethanol-treated antigen. Only one MAb that recognized site I reacted with the denatured G protein in a Western blotting assay, indicating that its epitope is linear. These results suggest that almost all of the epitopes on the G protein of the rabies virus are conformation-dependent and the G protein forms a complicated antigenic structure.

Antibodies, Monoclonal↗

Neural cell adhesion molecule L1 in gliomas: correlation with TGF-beta and p53.

AIMS: To assess immunohistochemically whether the neural cell adhesion molecule L1, which is a member of the immunoglobulin superfamily and has been shown recently to be a stimulating factor for glioma migration, is expressed in glioma tissues, and to investigate factors that can regulate this expression. METHODS: Twenty seven glioma tissue specimens including 13 glioblastomas, seven anaplastic astrocytomas, and seven astrocytomas were examined. Immunohistochemical analyses of L1, p53, and transforming growth cell factor beta (TGF-beta) were performed on each tumour using both polyclonal and monoclonal antibodies. RESULTS: Nine (33%) specimens (six glioblastomas and three anaplastic astrocytomas) had L1 positive immunostaining. p53 positive staining was detected in 10 (43%) of 23 glioma specimens (seven glioblastomas and three anaplastic astrocytomas). TGF-beta positive immunostaining was observed in 12 (52%) of the 23 glioma specimens (six glioblastomas, four anaplastic astrocytomas, and two astrocytomas). There was a statistical correlation between both p53 and L1 expression and TGF-beta and L1 expression. No such correlation was found between p53 and TGF-beta expression. CONCLUSIONS: These results suggest that mutation of the p53 gene or expression of TGF-beta may upregulate the expression of the L1 gene, thus resulting in high grade migration of glioma cells.

Antigens, Neoplasm↗

Seroprevalence of Bartonella henselae and Toxoplasma gondii infections among pet cats in Kanagawa and Saitama Prefectures.

Seroprevalence of Bartonella henselae and Toxoplasma gondii was investigated among 471 pet cats obtained from seven private animal hospitals in Kanagawa and Saitama Prefectures during the period from May 1994 to June 1995. 'Furthermore, 67 randomly selected from the 471 serum samples were examined for the feline immunodeficiency virus (FIV) antibody and feline leukemia virus (FeLV) antigen. The antibody to B. henselae was examined by an indirect immunofluorescent antibody test. T. gondii, FIV and FeLV infections in cats were detected with respective commercial kits. Of the cat serum samples tested, 43 (9.1%) were found to be seropositive for B. henselae and 41 (8.7%) for T. gondii. The B. henselae-positive rate (12.9%) of male cats was significantly higher than that (5.2%) of female cats. On the other hand, T. gondii-positive rate was 9.1% in male and 8.7% in female cats and there was no significant difference in the positivity between sexes. The positive rate in each hospital varied from 0 to 19.5% for B. henselae and 4.9 to 18.8% for T. gondii. The ages of B. henselae- and T. gondii-positive cats were distributed from < 1-year-old to 14-year-old and the seropositivity increased with age of cats. Of the 67 cat serum samples, 16 and 6 cases were positive for FIV and FeLV, respectively. There was no relationship between these viral and B. henselae infections in cats.

Animals↗

Subcellular distribution of actively partitioning F plasmid during the cell division cycle in E. coli.

F plasmid is partitioned with fidelity to daughter cells during cell division cycle owing to two trans-acting genes, sopA and sopB, and a cis-acting site, sopC. We visualized the subcellular distribution of mini-F-plasmid molecules by fluorescence in situ hybridization. Mini-F-plasmid molecules having the sopABC segment were localized at midcell in newborn cells. Replicated plasmid molecules migrated to cell positions 1/4 and 3/4 without coupling with cell elongation and were tethered to these positions until completion of cell division. In contrast, molecules of a mini F plasmid lacking the sopABC segment were distributed randomly in spaces not occupied by nucleoids. The sopABC system caused replicated plasmid molecules to be positioned and tethered at the cell quarter sites.

Cell Division↗

Microsatellite instability and mutated type II transforming growth factor-beta receptor gene in gliomas.

Microsatellite instability has been reported in familial cancer syndrome and in various kinds of human sporadic tumors. We investigated the replication error (RER) and mutation rate of the transforming growth factor-beta type II receptor (TGF-beta RII) gene to determine the frequency of the RER+ phenotype and elucidate the relation between the mutation of the TGF-beta RII gene and RER in the tumorigenesis of glioma. We screened genomic DNA from 40 gliomas, comprised from 24 glioblastomas (GB), 11 anaplastic astrocytomas (AA) and five astrocytomas (AS) and compared the results with DNA from corresponding leukocytes. Seven of the 40 (18%) gliomas had the RER+ phenotype: five (21%) of 24 GB and two (18%) of 11 AA. In six gliomas we detected mutation of the TGF-beta RII gene. Five (71%) of seven RER+ and one (3%) of 33 RER-tumors had one A deletion in the (A)10 repeat of the TGF-beta RII gene. No mutations were detected in the (GT)3 repeat area of the TGF-beta RII gene. As the normal cells of these glioma patients had no mutations, we concluded that the mutations were somatic. We posit that the observed mutations inactivate the receptor through a frameshift mutation resulting in protein truncation. Our data suggest that the TGF-beta RII (A)10 repeat may be one area of genomic instability in the early stages of malignant glioma tumorigenesis.

Astrocytoma↗

Acute disseminated demyelination due to primary human herpesvirus-6 infection.

UNLABELLED: A previously healthy 19-month-old boy developed acute encephalopathy, thrombocytopenia and hepatic dysfunction. Human herpesvirus-6 (HHV-6) DNA was found in his CSF during the acute stage of the disease by means of the polymerase chain reaction. T2-weighted MRI revealed high signal intensity in the left thalamus and left parieto-occipital deep white matter. The myelin basic protein concentration in the CSF was elevated suggesting acute demyelination. The patient is now 2.5 years old and has no sequelae. CONCLUSION: Since clinical course and neuroimaging after HHV-6 infection are similar to those in acute disseminated encephalomyelitis, clinicians must pay attention to primary HHV-6 infection in patients under 2 years old with white matter lesions.

Acute Disease↗

Sequence analysis of the VP6 gene in group A turkey and chicken rotaviruses.

cDNAs corresponding to the VP6 gene of the turkey rotavirus strains Ty-1 and Ty-3, and the chicken rotavirus strain Ch-1, were cloned and sequenced. The nucleotide and deduced amino acid sequence homology in the coding region of the VP6 gene in avian rotaviruses ranged from 78.1 to 93.9% and 86.1 to 98.7%, respectively. Both sequences of VP6 from avian rotaviruses exhibited a low degree of sequence homology (67.8-70.7% and 69.8-74.6%, respectively) compared with mammalian rotaviruses. Phylogenetic tree analysis showed that all avian rotaviruses were included in a single cluster and have separated early or from mammalian rotaviruses during evolution. The chicken rotavirus strain Ch-1 was a distant relative of other avian rotaviruses.

Amino Acid Sequence↗

A virus-neutralizing epitope on the glycoprotein of rabies virus that contains Trp251 is a linear epitope.

We have established a hybridoma producing monoclonal antibody (MAb) against a linear epitope of glycoprotein (G protein) of the RC-HL strain of rabies virus. This MAb15-13 showed almost the same neutralizing activity to all of five rabies fixed strains, including RC-HL, and reacted to the denatured G protein in western blot analysis. To characterize and map this linear epitope, an antigenic variant NR15-13 was selected from RC-HL strain in the presence of neutralizing MAb15-13. The variant reacted with MAb15-13 in an immunofluorescent antibody test but was not neutralized by the antibody and the antibody did not bind to the variant G protein in a Western blot analysis. The variant NR15-13 had an amino acid substitution at position 251 of the G protein, where tryptophan of the parental RC-HL strain was replaced by arginine. Site-directed mutagenesis analysis using the expression system in simian COS7 cells revealed that a single amino acid substitution at 251-tryptophan by arginine on the G protein of the parental RC-HL strain abolished the antigenicity of the epitope for MAb15-13 in western blot analysis, and the replacement of 251-arginine by tryptophan recovered the activity. These results strongly suggest that tryptophan at position 251 on the G protein is essential for construction of the linear epitope against MAb15-13.

Animals↗

Fibronectin-mediated cell migration promotes glioma cell invasion through chemokinetic activity.

In order to investigate the biological role of fibronectin in glioma cell invasion, we studied the relation between migratory responses or adhesiveness of glioma cells to fibronectin and the in vitro invasion in three human malignant glioma cell lines, A172, T98G and U373MG. All these cell lines chemotactically migrated in a dose-dependent manner to fibronectin in concentrations ranging from 0.5 to 10 microg/ml, with A172 cells showing the strongest migration and U373 cells the weakest. Checkerboard analyses demonstrated that A172 and T98G cells showed much stronger chemokinetic responses to fibronectin than U373MG cells. In contrast to the migratory responses, A172 and U373MG cells showed an almost equally high adhesion to fibronectin and T98G cells a low adhesion. The degree of expression of the integrin alpha5 subunit correlated well with the strength of glioma cell adhesion to fibronectin rather than that of migration to the molecule. Furthermore, the cell adhesion to fibronectin was almost completely inhibited by arginine-glycine-aspartic acid (RGD)-containing peptides, but the fibronectin-stimulated cell migration was only partially inhibited. An in vitro invasion assay disclosed that U373MG cells invaded the artificial basement membrane barrier the most and A172 cells the least. However, addition of fibronectin to the glioma cells markedly enhanced the invasive activity of A172 and T98G cells but had little effect on that of U373MG cells. These results indicate that fibronectin-stimulated migration can be one of the factors promoting invasiveness of glioma cells and that the chemokinetic activity of fibronectin may play a crucial role in glioma invasion through conferring motor-driving force on the glioma cells.

Cell Adhesion↗

Focal encephalitis mimicking malignant lymphoma of the central nervous system.

We present a case of subacute focal encephalitis in a 62-year-old man whose clinical manifestations, neuroradiological findings and histopathological features mimicked those of a patient with malignant lymphoma of the central nervous system (CNS). In addition to routine histopathological examination, extensive studies including serological examination for encephalitic viruses, immunohistochemistry and DNA analysis were required to distinguish focal encephalitis from lymphoma of the CNS, clinical entities which differ in treatment.

Acyclovir↗

Functional evaluation of the remaining kidney in kidney donors by radionuclide dynamic imaging using a graphic method with factor analysis.

An uptake coefficient proportional to the glomerular filtration rate (GFR) can be estimated from dynamic renal images with 99Tcm-DTPA using Rutland's graphic method. We have developed a new method of extracting the input and retention functions by applying factor analysis to the renal dynamic images obtained with 99Tcm-DTPA, which we have called the 'factor uptake coefficient' (Factor UC). In the present study, we followed 13 living kidney donors (7 males, 6 females) by measuring the Factor UC in each kidney before nephrectomy and in the remaining kidney 3 weeks and 1 year after nephrectomy. The median Factor UC in the remaining kidney increased from a pre-nephrectomy value of 0.31 to 0.52 three weeks post-nephrectomy, which then remained unchanged for up to 1 year. These results indicate that functional compensation occurs following unilateral nephrectomy, and that this process is complete within 3 weeks after nephrectomy.

Adult↗

A new competitive enzyme-linked immunosorbent assay demonstrates adequate immune levels to rabies virus in compulsorily vaccinated Japanese domestic dogs.

A competitive enzyme-linked immunosorbent assay (c-ELISA) was developed as an alternative to the viral neutralization (VN) test for rapid and simple detection of antibodies to rabies virus. The competitor antibody in the c-ELISA was a biotinylated monoclonal antibody to the nucleoprotein of rabies virus. Initial comparisons showed a high correlation between titers obtained with the VN test and the c-ELISA (n = 88, r = 0.90), indicating that the c-ELISA could be used as a reliable substitute for the VN test. To evaluate the immune status of Japanese dogs to rabies virus, a total of 1,019 serum samples were collected from domestic dogs in 1994 and tested for antibodies with the c-ELISA. Overall, 84.8% of the dogs had antibodies against rabies virus, indicating that the vaccination strategy for preventing rabies outbreaks in domestic dogs is probably sufficient in Japan. Dogs receiving final vaccinations a year or more previously were 48.3 and 90.3% positive for antibodies when vaccinated once only or two or more times, respectively. This suggests that almost all dogs vaccinated twice or more remain seropositive for over 1 year in Japan.

Animals↗

Cellular proliferation and secretion in secondary hyperparathyroidism during renal failure.

Secondary hyperparathyroidism is one of the severe complications of chronic renal failure. In this study, we investigated the cellular components of parathyroid tissue, with measurements of various serum parathyroid hormone (PTH) types in the circulation, and evaluated their clinical significance in hemodialysis patients. Thirty-eight patients who underwent both subtotal parathyroidectomy and autotransplantation in Tokai University Hospital from 1979 to 1994 were divided into two groups. Group 1 (G-1) was not administered active-type vitamin D3 (VD; 19 patients; 15 males and 4 females), and group 2 (G-2) was administered VD (19 patients; 10 males and 9 females). The parathyroid tissues which were obtained from G-1 and G-2 patients were classified into three classes based on the size of the oxyphilic cell area (class I < 25%, class II 25-50% and class III > 50%). Our results showed that the proliferation index of parathyroid tissues in the oxyphilic cell area was higher in G-2 than that in G-1. Immunohistochemically, MIB-1 staining was more intense than that in the chief cell area in G-2 patients. Moreover, the proliferative index in the same specimen was also higher in the oxyphilic cell area than in the chief cell area. It was suggested that the oxyphilic cells proliferated independently. Furthermore, synthesis of PTH in the oxyphilic cell area was revealed immunohistochemically by the presence of PTH and was confirmed by positive staining of PTH mRNA in the oxyphilic cell area with in situ hybridization. HS(M-terminal)- and C-PTH levels in the serum were significantly higher in class III than in class I (p < 0.01). No significant difference of HS- and C-PTH levels between class II and class III was noted. Moreover, no significant difference of intact-PTH levels was found in all three classes. From the above findings, it was suggested that proliferation of the parathyroid tissues or secretory state of PTH in hemodialysis-maintained patients with secondary hyperparathyroidism, which was closely related to the proliferation of oxyphilic cells, can be more accurately reflected by HS- and C-PTH levels than by the intact-PTH level. Therefore, it was suggested that HS- and C-PTH levels in the serum are important indices for accurate evaluation of the pathology and suitable therapy of secondary hyperparathyroidism, as well as observation of the clinical course.

Adult↗